1.Characterization of human αβTCR repertoire and discovery of D-D fusion in TCRβ chains.
Peipei LIU ; Di LIU ; Xi YANG ; Jing GAO ; Yan CHEN ; Xue XIAO ; Fei LIU ; Jing ZOU ; Jun WU ; Juncai MA ; Fangqing ZHAO ; Xuyu ZHOU ; George F GAO ; Baoli ZHU
Protein & Cell 2014;5(8):603-615
The characterization of the human T-cell receptor (TCR) repertoire has made remarkable progress, with most of the work focusing on the TCRβ chains. Here, we analyzed the diversity and complexity of both the TCRα and TCRβ repertoires of three healthy donors. We found that the diversity of the TCRα repertoire is higher than that of the TCRβ repertoire, whereas the usages of the V and J genes tended to be preferential with similar TRAV and TRAJ patterns in all three donors. The V-J pairings, like the V and J gene usages, were slightly preferential. We also found that the TRDV1 gene rearranges with the majority of TRAJ genes, suggesting that TRDV1 is a shared TRAV/DV gene (TRAV42/DV1). Moreover, we uncovered the presence of tandem TRBD (TRB D gene) usage in ~2% of the productive human TCRβ CDR3 sequences.
Complementarity Determining Regions
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genetics
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DNA Primers
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chemistry
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genetics
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Female
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Gene Rearrangement, beta-Chain T-Cell Antigen Receptor
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genetics
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Gene Rearrangement, delta-Chain T-Cell Antigen Receptor
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genetics
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Genes, T-Cell Receptor beta
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genetics
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Genetic Variation
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High-Throughput Nucleotide Sequencing
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Humans
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Immunoglobulin Joining Region
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genetics
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Immunoglobulin Variable Region
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genetics
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Male
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Receptors, Antigen, T-Cell, alpha-beta
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genetics
2.Molecular Diagnosis of Cutaneous T Cell Lymphoproliferative Diseases.
Ji Young PARK ; Myung Hoon LEE ; Eun Kyung KWAK ; Dong Ja KIM ; Tae In PARK ; Han Ik BAE
Korean Journal of Pathology 2000;34(11):941-949
It is often problematic to diagnose T-cell lymphoproliferative disorders of the skin because of the difficulty in establishing clonality in paraffin-embedded tissue. We used polymerase chain reaction single strand conformational polymorphism (PCR-SSCP) and heteroduplex analysis in paraffin embedded tissue to detect clonal rearrangement of T-cell receptor gamma (TCRgamma) gene in 17 T-cell lymphoproliferative disorders and 6 atypical lymphoproliferative diseases. We used polymerase chain reaction to detect TCR beta gene rearrangement in 8 of 17 cases which did not show TCRgamma gene rearrangement. Jurkat cell lines were used as monoclonal controls. DNA was extracted from 5 biopsies of T-cell lymphomas, 10 biopsies of mycosis fungoides, 2 biopsies of lymphomatoid papulosis, and 6 biopsies of atypical lymphoproliferative lesions. We detected monoclonality in 5 of 5 T-cell lymphoma cases, 2 of 2 lymphomatoid papulosis cases, 6 of 10 mycosis fungoides cases, and 2 of 6 atypical lymphoproliferative disease cases. We conclude that nonradioactive PCR-SSCP for TCR gene rearrangement analysis is a useful adjunct to routine histological and immunophenotypic methods in the diagnosis of cutaneous T cell lymphoproliferative disorders in paraffin embedded tissue.
Biopsy
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Diagnosis*
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DNA
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Gene Rearrangement
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Genes, T-Cell Receptor
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Genes, T-Cell Receptor beta
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Heteroduplex Analysis
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Humans
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Jurkat Cells
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Lymphoma, T-Cell
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Lymphomatoid Papulosis
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Lymphoproliferative Disorders
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Mycosis Fungoides
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Paraffin
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Polymerase Chain Reaction
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Receptors, Antigen, T-Cell
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Skin
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T-Lymphocytes
3.Characteristics of T cell receptors recognizing antiphospholipid syndrome associated antigens.
Yuan OU ; Ping ZHU ; Xia ZHU ; Jiang-Ying GU ; Jing LIU ; Jin-Wei DU ; Ying ZHANG ; Hong-Xing LIU ; Xin ZHUANG
Journal of Experimental Hematology 2006;14(6):1156-1159
To understand the characteristics of T cell receptors recognizing antiphospholipid syndrome associated antigen, the characteristics of T cells were analyzed using T cell receptor beta variable region (TCRbetaV) gene spectrotyping in a case of antiphospholipid syndrome (APS). The results indicated that in the case of APS there were 2 dominant T cell clones. The TCRbetaVs sequences of the 2 T cell clones showed the TCRbetaVs belonged to 8 and 23 gene families respectively. The peptides of third complementarity-determining regions (CDR3) in the TCRbetaVs were CASSLLVAGGPRAYNEQFFGPG and CASSLAGFGQPQHFGDG. Comparing the motifs in CDR3 with another autoimmune disease, the motif YNEQFFGPG in TCRbetaV8 and motif QHFGDG in TCRbetaV23 were identical with that of idiopathic thrombocytopenic purpura and systemic lupus erythematosus reported before. In conclusion, some T cell clones proliferating in these autoimmune diseases may recognize the same antigens.
Adult
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Antiphospholipid Syndrome
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immunology
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Autoantigens
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immunology
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Female
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Gene Rearrangement, alpha-Chain T-Cell Antigen Receptor
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immunology
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Genes, T-Cell Receptor beta
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genetics
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immunology
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Humans
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Immunoglobulin Variable Region
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immunology
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Receptors, Antigen, T-Cell
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immunology
4.Analysis of T lymphocytic clones in diseases by spectrotyping of T cell beta variable region - review.
Xia ZHU ; Xiao-Ling GUO ; Ping ZHU
Journal of Experimental Hematology 2005;13(4):703-708
T cells recognize antigens through TCRs (T cell receptor). T cell clones can be sorted into 24 gene subfamilies based on the usage of the segments of TCR BV in gene rearrangement. Application of the various segments of TCR BV may establish TCR BV spectrotyping that can be used to analyze and recognize the different functional T cell clones, and understand the function and proliferation of various T cell clones in malignancy and autoimmune disease. In vitro expansion of a great deal of the specific antitumor T cells and transfusing them to patients will be able to develop a new method for tumor immunotherapy. Through analyze the character of the TCR BV gene, McAb against TCR or DNA vaccine to inhibit the growth of T cell clones associated-autoimmune disease and tumors might be developed. The McAb and vaccine may be used to cure these diseases. The commo T cells can also be modifed to specific antitumor T cells by method of TCR gene transfer. In this review, the characteristics of TCR, analysis method for gene spectrotyping of TCR BV, segments of TCR BV and autoimmue distase, T cell clones in hematologic maligrancies, recognition of T cell oligoclone expansion of T cells, and application of TCR BV gene spectrotyping in bone marrow transplantation were discussed and summarised.
Autoimmune Diseases
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genetics
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immunology
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Clone Cells
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cytology
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metabolism
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Gene Rearrangement, beta-Chain T-Cell Antigen Receptor
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genetics
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Hematologic Neoplasms
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genetics
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immunology
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Humans
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Receptors, Antigen, T-Cell
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classification
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genetics
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T-Lymphocyte Subsets
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cytology
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metabolism
5.Analysis of T cell receptor BV dominant usage and CDR3 sequences during acute exacerbation in patients with chronic hepatitis B.
Guang-wen ZHANG ; Xin-sheng YAO ; Shi-wu MA ; Chuang-guo YANG ; Yue-cheng YU ; Jin-lin HOU
Chinese Journal of Hepatology 2006;14(1):23-28
OBJECTIVESTo understand the role cellular immunology plays in the pathogenesis of chronic hepatitis B (CHB) through analysis of T cell receptor (TCR) beta chain variable region gene (BV) family dominant usage and beta chain complementarity determining region3 (CDR3) sequences of peripheral blood mononuclear cells of the patients.
METHODSTCR BV families were amplified by inverse polymerase chain reaction (RT-PCR), and the dominant usage of BV families and CDR3 repertoire were analyzed by immunoscope technology for 8 CHB patients during their acute exacerbations and for 4 healthy blood donors who served as controls. The clonality of the T cells suspected by immunoscope was further confirmed by CDR3 sequencing.
RESULTSThe TCR BV CDR3 repertoire of the 4 healthy blood donors showed a Gaussian distribution. In the 8 CHB patients, however, the clonal expansion of T cells showed different TCR BV families with each patient. The T cells of the clonal expansion shared different CDR3 sequences.
CONCLUSIONThe peripheral blood T cells of CHB patients during their acute exacerbation showed significantly a clonal expansion and their T cell clonal expansion may be stimulated by several HBV epitopes. These results indicate that cellular immunology is involved in the pathogenesis of the liver inflammation process of CHB.
Adult ; Amino Acid Sequence ; Base Sequence ; Cloning, Molecular ; Complementarity Determining Regions ; genetics ; Female ; Gene Rearrangement, beta-Chain T-Cell Antigen Receptor ; genetics ; Hepatitis B, Chronic ; genetics ; Humans ; Male ; Middle Aged ; Molecular Sequence Data ; Receptors, Antigen, T-Cell ; genetics
6.Significance of TCR gene clonal rearrangement analysis in diagnosis of mycosis fungoides.
Chen XU ; Yuan TANG ; Lin WANG ; Chuan WAN ; Wei-ping LIU
Chinese Journal of Oncology 2010;32(9):685-689
OBJECTIVETo investigate the significance of detecting TCR gene clonal rearrangement in the diagnosis of mycosis fungoides (MF) and to optimize the primers used for detecting the TCR gene clonal rearrangement with PCR in paraffin embedded tissues of MF.
METHODSNineteen cases of MF were enrolled into the study. A panel of 10 antibodies were used for immunophenotypic analysis and polymerase chain reaction for TCR-γ and TCR-β gene rearrangement detection in this study.
RESULTSTCR gene clonal rearrangements were detected in all 19 cases, in which 84.2% cases (16/19) had TCR-γ gene clonal rearrangements. The positive rates of the primers T(VG)/T(JX), V(2-5)/V(8-12)/JGT(1) and BIOMED-2-TCR-γ were 47.4%, 78.9% and 31.6%, respectively. The positive rate of V(2-5)/V(8-12)/JGT(1) was statistically significantly higher than that of T(VG)/T(JX) and BIOMED-2-TCR-γ (P < 0.05). No TCR gene clonal rearrangement was detected using the primers V(γ11)/V(γ101)/Jγ12 and V(γ11)/V(γ101)/J(p12). TCR-β gene clonal rearrangement was detected in 31.6% (6/19) cases.
CONCLUSIONSTCR gene clonal rearrangement analysis is a useful tool in the diagnosis of MF and TCR-γ gene is a good target gene for the detection. The primers T(VG)/T(JX), V(2-5)/V(8-12)/JGT(1) and BIOMED-2-TCR-γ can be used in clinicopathologic detection for TCR gene clonal rearrangement and V(2-5)/V(8-12)/JGT(1) may be the first choice.
Adolescent ; Adult ; Aged ; Antigens, CD7 ; metabolism ; Base Sequence ; CD2 Antigens ; metabolism ; CD3 Complex ; metabolism ; CD4 Antigens ; metabolism ; Child ; Child, Preschool ; Female ; Gene Rearrangement, beta-Chain T-Cell Antigen Receptor ; Gene Rearrangement, gamma-Chain T-Cell Antigen Receptor ; Humans ; Leukocyte Common Antigens ; metabolism ; Male ; Middle Aged ; Molecular Sequence Data ; Mycosis Fungoides ; diagnosis ; genetics ; metabolism ; pathology ; Paraffin Embedding ; Receptors, Antigen, T-Cell, alpha-beta ; genetics ; Receptors, Antigen, T-Cell, gamma-delta ; genetics ; Skin Neoplasms ; diagnosis ; genetics ; metabolism ; pathology ; Young Adult
7.Application of BIOMED-2 primers in analysis of T-cell receptor gamma gene rearrangements in paraffin-embedded tissue specimens of T-cell lymphoma.
Yuan TANG ; Wei JIANG ; Lei LI ; Hong JI ; Yun LI ; Gan-di LI ; Wei-ping LIU
Chinese Journal of Pathology 2009;38(4):253-257
OBJECTIVETo evaluate the practical values of PCR detectable T-cell receptor (TCR) gene rearrangement in paraffin embedded tissue samples in the diagnosis of T-cell malignancies using BIOMED-2 PCR multiplex tubes TCRgamma(A+B).
METHODSTraditional phenol-chloroform method was used to extract DNA from 55 cases of archival paraffin embedded tissues samples of T-cell malignancies and the DNA quality was evaluated by PCR-based amplification of housekeeping gene beta-globin. The selected BIOMED-2 PCR multiplex tubes TCRgamma(A+B) were used to detect TCR gene rearrangement and comparison with the results of universal TCR primers (T(VG)/T(JX)) was performed.
RESULTSPositive detection rates by the BIOMED-2 multiplex tubes TCRgamma(A+B) and the universal primers (T(VG)/T(JX)) were 76.4% and 60.0%, respectively. There were not statistical difference between the methods (P > 0.05).
CONCLUSIONBIOMED-2 multiplex tubes TCRgamma(A+B) is suitable for detection of clonal rearrangements of TCR genes in current archival paraffin embedded tissue samples of T-cell malignancies.
DNA Primers ; DNA, Neoplasm ; analysis ; Gene Rearrangement, gamma-Chain T-Cell Antigen Receptor ; Humans ; Lymphoma, T-Cell ; genetics ; metabolism ; pathology ; Paraffin Embedding ; Polymerase Chain Reaction ; methods ; beta-Globins ; metabolism
8.Diagnostic significance of TCR gene clonal rearrangement analysis in early mycosis fungoides.
Chen XU ; Chuan WAN ; Lin WANG ; Han-Jun YANG ; Yuan TANG ; Wei-Ping LIU
Chinese Journal of Cancer 2011;30(4):264-272
Mycosis fungoides (MF), the most common type of cutaneous T-cell lymphoma, has various unspecific clinical and histological characteristics. Its early diagnosis is challenging. The application of T-cell receptor (TCR) gene clonal rearrangement to the diagnosis of MF has been widely studied. In this study, we used polymerase chain reaction (PCR) to investigate the diagnostic significance of detecting TCR-γ and -β gene clonal rearrangement in the early diagnosis of mycosis fungoides. PCR for TCR-γ and TCR-β gene rearrangement was performed on 19 patients with suspected early MF, 6 with typical MF, and 6 with chronic dermatitis. Of the 19 patients with suspected early MF, 13 had TCR-γ gene clonal rearrangement, whereas none had TCR-β gene clonal rearrangement. All patients with typical MF had TCR gene clonal rearrangement, in which 4 showed TCR-γ clonal rearrangement, 1 showed TCR-β gene clonal rearrangements, and 1 showed both. No patients with chronic dermatitis had TCR gene clonal rearrangement. These results indicate that TCR gene clonal rearrangement analysis is a useful tool in diagnosing early MF. TCR-γ gene is recommended to the routine analysis, whereas TCR-β gene has potential in combination toward intractable cases.
Adolescent
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Adult
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Aged
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Base Sequence
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genetics
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DNA, Neoplasm
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genetics
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Early Detection of Cancer
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methods
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Female
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Gene Rearrangement, beta-Chain T-Cell Antigen Receptor
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Gene Rearrangement, gamma-Chain T-Cell Antigen Receptor
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Humans
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Male
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Middle Aged
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Mycosis Fungoides
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diagnosis
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genetics
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Polymerase Chain Reaction
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Skin Neoplasms
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diagnosis
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genetics
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Young Adult
9.Changes in T-cell receptor repertoire in aplastic anemia and effects of Shengxue Mixture.
Yong-ming ZHOU ; Xue-li WEI ; Jia-hui LU
Chinese Journal of Integrated Traditional and Western Medicine 2006;26(11):973-977
OBJECTIVETo explore the immune pathogenesis of aplastic anemia (AA) and the therapeutic effects of Shengxue Mixture (SM) through the gene expressions of subfamilies of T-cell receptor variable region beta (TCR Vbeta) using immunologic and molecular biologic technology.
METHODSGene expressions of TCR Vbeta sub-families in peripheral blood mononuclear cells from 20 AA patients were detected before and after treatment with SM using RT-PCR and gene scanning method.
RESULTSTCR Vbeta gene repertoire of the 24 subfamily genes deviated in AA patients, and the oligoclonal gene expressions increased obviously compared with those in healthy people (P < 0.01), including Vbeta2, 5, 6, 15, 16, 22, and 23 were found in 30%-50% AA patients, and Vbeta8, 21 were in more than 50% patients. These oligoclonal genes reduced significantly after treatment with SM compared with those before treatment (P < 0.05).
CONCLUSIONMultiple TCR Vbeta subfamilies of clonal proliferation participate in the pathogenesis of AA. SM can rectify the deviation of TCR Vbeta gene repertoire, reduce the abnormal clonal proliferation of T cells, thus to alleviate the immune injury to hematopoietic tissue, and thus to benefit the recovery of hematopoiesis of bone marrow.
Adolescent ; Adult ; Aged ; Anemia, Aplastic ; drug therapy ; genetics ; immunology ; Drugs, Chinese Herbal ; pharmacology ; therapeutic use ; Female ; Gene Expression ; drug effects ; Gene Rearrangement, beta-Chain T-Cell Antigen Receptor ; Humans ; Male ; Middle Aged ; Phytotherapy ; Receptors, Antigen, T-Cell, alpha-beta ; biosynthesis ; genetics ; Reverse Transcriptase Polymerase Chain Reaction
10.Effects of T cell receptor gene rearrangement on BV CDR3 in Jurkat cells.
Hong-Yun ZOU ; Li MA ; Xin-sheng YAO ; Qian WEN ; Wei LUO ; Xiao-Ning WANG
Journal of Southern Medical University 2006;26(7):939-943
OBJECTIVETo investigate the effects of T cell receptor (TCR) BD2-BJ2 gene rearrangement on the complementary-determining region (CDR) 3 of TCR beta chain (TCR BV CDR3) in Jurkat cells.
METHODSTCR BV gene subfamilies were detected by RT-PCR in Jurkat cells during proliferation and after induction with non-specific T cell activators and SEA, respectively. To determine the clonality of TCR BV subfamilies and the lengths of CDR3, the PCR products were analyzed by TCR GeneScan technique, and the sequences of CDR3 were further analyzed by DNA sequencer.
RESULTSNo new TCR BV subfamilies were found in Jurkat cells, a monoclonal BV8(+)cell line, either during cell proliferation or after stimulation with different treatments, nor were any differences found in CDR3 size or sequences.
CONCLUSIONTCR BD2-BJ2 rearrangement in Jurkat cells may not play a role in modification of TCR BV CDR3 domains or the consequent antigen immunorecognition of BV CDR3, but the possibility of TCR modification can not be excluded.
Amino Acid Sequence ; Base Sequence ; Complementarity Determining Regions ; genetics ; Gene Rearrangement, beta-Chain T-Cell Antigen Receptor ; genetics ; Humans ; Immunologic Factors ; genetics ; Jurkat Cells ; Leukemia, T-Cell ; genetics ; immunology ; pathology ; Molecular Sequence Data ; Receptors, Antigen, T-Cell ; genetics ; T-Lymphocytes ; immunology ; metabolism